User:Anthony Salvagno/Notebook/Research/2010/10/19/Tethering To-Do
With the tweezer ready for tweezing, calibration under-way, and some last minute mechanical features being added it's time to get tethering, stretching, and unzipping going again. I need to get my head together to get it working at least as efficiently as it was before BPS last year. Yesterday I did some dabbling and it wasn't very good. Things need to be better.
- make new BGB
- I have a 50ml tube of it ready to go. It just needs to be filtered. I made this last June/July I think. Is it ok to go?SJK 21:10, 19 October 2010 (EDT)
- make new POP
- I definitely need new pop and I think I need to remake one chemical in the mix.
- prepare more anti-dig SJK 21:11, 19 October 2010 (EDT)
- Get new beads
- small beads, the big beads are fine except they don't tether well
- try cleaning with Windex
- may be something magical in the windex that gave Koch great tethering. This is literally the only thing I've yet to do from Koch's glory days.
- SJK 21:12, 19 October 2010 (EDT)
- Right now I'm using 1:100 concentration for the 1.1kb tethers I think this puts it somewhere in the 10-100pM range
- I need to mix my DNA tubes up. They've been sitting in the lab top coolers for some time now.
- For DNA I have: TpBR (tetherable pBR fragments), Tyeast (I'm calling it that for now, it's some kind of shotgun clone), 1.1kb dsDNA tethers, 4.4kb dsDNA tethers (11pM I believe)
- Since I'm starting with the dsDNA tethers (for stretching) should I use my frozen stock or will mixing it up do the trick? I guess I will try mixing first (gently of course) so I don't have to throw anything away. SJK 21:13, 19 October 2010 (EDT)